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Using single-nucleus RNA-sequencing to interrogate transcriptomic profiles of archived human pancreatic islets

ABSTRACT: Human pancreatic islets, including insulin secreting beta-cells are a major focus of transplantation strategies aimed at identifying new therapeutic approaches to counteract hyperglycemia in patients with diabetes. Identifying the transcriptomic signature of human islet cells provides insights into regulatory pathways that can be harnessed for planning therapeutic strategies. In this context, single-cell RNA-sequencing (scRNA-seq) has been used mostly in vitro. However, in experimental human islet transplantation models the small amount of tissue is principally used for immunostaining and poses a challenge in performing ‘omics’ studies that provide unbiased information. To circumvent this limitation, we report the use of single nucleus RNA-sequencing (snRNA-seq) on frozen/archived human islet grafts, to define the transcriptomic signature of islet cells preserved after in vivo studies. Interrogating nuclear RNA, we were able to successfully identify all islet endocrine cells, obtain adequate coverage of genes and define molecular pathways that are important for studying human islet cell biology (e.g. cell cycle, apoptosis, insulin secretion). Intersecting our nuclear transcriptomic output with publicly available single-cell RNA-seq datasets, revealed ~90% overlap of the detected genes. In conclusion, we propose that snRNA-seq represents a reliable strategy to probe transcriptomic profiles of fresh or archived transplanted human islets.

ORGANISM(S): Homo sapiens

PROVIDER: GSE150212 | GEO | 2021/07/09

REPOSITORIES: GEO

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